Gastric Submucosal Tissue as a Novel Diagnostic Tool

Inactive Publication Date: 2010-02-18
PURDUE RES FOUND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0011]The present invention is directed to an extracellular matrix, comprising vertebrate submucosal tissues, that provides the necessary microenvironment to allow the in vitro culture of fastidious organisms. Naturally occurring extracellular matrices have the ability to serve as a substrate for the growth of prokaryotic organisms, fungal agents and less well defined infectious agents of different genus and species which are difficult to grow in standard culture media. The extracellular matrixes of the present

Problems solved by technology

However, many detected strains have proven to be difficult to culture outside their natural micronenvironment due to the unique culture conditions required by these organisms.
Many infectious agents when placed on existing culture media often fail to grow and therefore are not detected, given the state of the present technology.
Many bacteria cannot survive in an acidic environment, however H. pylori are not the only bacteria capable of colonizing the surface of a primate's stomach.
Although blood tests are useful for an initial screen for detecting the existence of an H. pylori infection, The blood test is based on detecting antibodies to

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of Stomach Submucosal Tissue

[0033]The tissue graft material of this invention is prepared in accordance with the following steps:

[0034]The stomach is first removed from the animal source by cutting the esophagus and small intestine at their respective entrance and exit points on the stomach. Any excess mesentery tissue or fat is removed from the stomach and the contents of the stomach are emptied and any remaining residues are removed from the inside of the stomach by rinsing with running tap water.

[0035]The stomach is then everted to expose the inside layers of the stomach. The portions of the stomach that begin to form the entrance or exit points of the stomach are removed. The stomach is typically left whole, however the stomach can also be cut and flattened prior to removal of unwanted tissues. The luminal surface of the stomach is subject to abrasion using the handle portion of a pair of scissors or hemostats to scrape off the inner layers of the stomach including a...

example 2

In-Vitro Cell Growth Properties of Stomach Submucosa

[0037]The ability of stomach submucosa to serve as an extracellular matrix to support in-vitro cell growth was tested by applying several cell types to the stomach submucosal tissue surface under standard cell culture conditions. The cell types tested included 3T3 fibroblasts, intestinal epithelium cells, and FR (fetal rat) mesenchymal cells. All three cell types showed the ability to proliferate readily upon this extracellular matrix without the addition of the supplements that would be needed to grow these cells on a plastic surface. Therefore, it can be concluded that the material contains necessary structure and composition “nutrients” to serve as a cell culture substrate for supporting cell growth.

example 3

Kirby-Bauer Test

[0038]To determine if stomach submucosa inhibits the growth of H. pylori, a Kirby-Bauer test was conducted. Individual colonies of H. pylori were isolated from a chocolate agar plate and used to inoculate a 1 ml. solution of sterile saline in a small tube. This sterile saline solution was then used to inoculate a chocolate agar plate through the use of a sterile cotton swab. A small piece of stomach submucosal tissue (approximately 25-50 mm. in diameter) was placed in the middle of the inoculated chocolate agar plate and pressed onto the surface of the plate to assure that the submucosa tissue sticks to the chocolate agar. The experiment was conducted in duplicate; two plates having the luminal side of the submucosa tissue in contact with the chocolate agar, and two plates having the abluminal surface of the submucosa tissue in contact with the agar. The plates were then incubated in a Campy jar in the 37° C. aerobic incubator for 3-4 days. After incubation the plate...

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PUM

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Abstract

A cell culture growth substrate comprising submucosal tissue of a warm-blooded vertebrate and a method for culturing fastidious organisms is described. Submucosal tissue used in accordance with the present invention supports the proliferation of cells when said cells are contacted with submucosal tissue under conditions conducive to cell proliferation.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation of U.S. application Ser. No. 09 / 319,841 filed Jun. 10, 1999, which is a U.S. national application of international application Serial No. PCT / US97 / 22729 filed Dec. 10, 1997, which claims priority to U.S. provisional application Ser. No. 60 / 032,686 filed Dec. 10, 1996.FIELD OF THE INVENTION[0002]The present invention relates to a submucosal tissue composition and the use of those compositions to promote growth of fastidious cells. More particularly, the present invention is directed to the use of submucosal tissue cell culture substrates to enhance the in vitro growth, and thus the identification and diagnosis, of vertebrate infections agents.BACKGROUND OF THE INVENTION[0003]Bacteria are a diverse group of organisms that live in a broad variety of environments. In particular, many bacterial species live both on and in vertebrate hosts. Bacteria that colonize the interior spaces of vertebrates typically ex...

Claims

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Application Information

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IPC IPC(8): C12N5/07C12N1/20C12N5/00G01N33/50G01N33/569
CPCC12N1/20C12N5/0068Y10S435/851G01N33/5091G01N33/56922C12N2533/92
Inventor BADYLAK, STEPHEN F.BODER, GEORGE B.VOYTIK-HARBIN, SHERRY L.WANINGER, MATTHEW S.BRIGHTMAN, ANDREW O.
Owner PURDUE RES FOUND INC
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